Preparation of Antibacterial Epidermal Skin Graft

1.应用抗菌肽提呈皮肤片研究了经sapecin基因修饰的表皮细胞的抗菌活性。将胚胎干细胞移植到小鼠体内后，E. coli或S.金黄色葡萄球菌感染。此外，我们构建了一个新的基因，它具有sapecin基因，含有凝血酶消化基序的接头和血小板生长因子（血小板生长因子）受体。在通过脂质体转染法转染后，将细胞用作为标记物的Myc标签的荧光抗体染色。然而，没有观察到抗菌活性，因为肽产量非常低。然后，制备了能够锚在细胞表面的BAM-抗菌肽，并对其抗菌活性进行了评价。将BAM-抗菌肽加入到小鼠成纤维细胞培养液中，几分钟后抗菌肽就能显示在细胞表面。当大肠杆菌。将培养液与该细胞接触，抗菌肽逐渐释放。另一方面，BAM-抗菌肽易于锚定在小鼠皮肤上，并且还观察到对E. coli.2.培养皮肤细胞片制备方法的建立通过明胶与转氨酶（TGase）交联制备可生物降解的水凝胶。结果表明，5 wt 5明胶和1U/ml TGase对NIH/3 T3成纤维细胞的增殖最有利。明胶凝胶中加入细胞粘附因子可促进细胞增殖。将合成肽RGDLLQ添加到明胶溶液中，其中LLQ基序凭借谷氨酰胺残基是TGase的底物。这些结果表明，TGase介导的细胞粘附因子掺入明胶基质增强细胞增殖，这种新的生物材料是一种有效的工具，伤口敷料或组织工程。

1. Application antibacterial peptide-presenting skin sheetAntibacterial activity of epitherial cell which was genetically modified with sapecin gene was examined. After transplanting of eptherial cells into mouse, E. coli or S. aureus was infected. Moreover, we constructed new gene which had sapecin gene, linker containing a thrombin digesting motif and the PDGF (blood platelet growth factor) receptor. After transfection by lipofection method, the cells were dyed with the fluorescence antibody to the Myc tag which is a marker. However, the antibacterial activity was not observed, because peptide production was very low. Then, BAM-antibacterial peptide which was able to anchor on the cell surface was prepared, and its antibacterial activity was evaluated. When BAM-antibacterial peptide was added to the culture medium of mouse fibroblast cell, antibacterial peptide was able to be shown to the cell surface in several minutes. When E coli. culture medium was contacted into this cell, antibacterial peptide was released gradually and. On the other hand, BAM-antibacterial peptide was easily anchored on mouse outer skin and also antibacterial activity was observed against E. coli.2. Establishment of the preparation method of cultured skin cell sheetThe biodegradable hydrogels were prepared though crosslinking of gelatin with transglutaminase (TGase). We found that the concentration of 5wt5 gelatin and 1 unit/ml TGase were optimum for the proliferation of NIH/3T3 fibroblast. The cell proliferation was enhanced by incorporation of cell adhesion factors in to gelatin hydorgels. Synthetic peptide, RGDLLQ, were added to the gelatin solution where LLQ motif is a substrate of TGase by virtue of a glutamine residue. These results suggest that TGase-mediated incorporation of cell adhesion factors into gelatin matrices enhanced cell proliferation and this novel biomaterial is a potent tool for wound dressing or tissue engineering.